THE APICOMPLEXAN

Overview of classification of parasites

Kingdom- Animalia

Subkingdom Protozoa Subkingdom Metazoa

phylum sarcomastigophora phylum nematoda
phylum sporozoa phylum platyhelminth
 phylum ciliophora
phylum microspora

 Kingdom  phylum  class  order  family  genus 

Characteristics of apicomplexa
 are microscopic, spore-forming intracellular Single-
celled organisms that multiply in human host
 have complex life cycle, well-developed asexual &
sexual stages.
 motility is absent in most cells except male gametes.
Apicomplexa....
 Sporozoans produce special spore like cells called
sporozoites ff sexual reproduction w/c are important in
transmission of infections;
 most form thick-walled zygotes called oocysts;
 Reproduction
 asexually by schizogony (merogony)
 sexually by sporogony
Two groups


 Blood and tissue coccidia:

 Plasmodium species & Babestia spp.
 Toxoplasma gondii
 Intestinal coccidia:

 Isospora belli

 Cryptosporidium parvum

 Cyclospora cayetanensis

most coccidia are particularly important as pathogens

in immunosuppressive conditions
Toxoplasmosis
Toxoplasmosis
Toxoplasmosis

Introduction
 Toxoplasma gondii is an animal coccidian
parasite which causes toxoplasmosis
 congenital toxoplasmosis the most serious
form of human infection.
 T. gondii has a worldwide distribution.
 Toxoplasma gondii is one of the most well studied
parasites because of its medical and veterinary
importance.
 1908-Nicolle and Manceaux
 It is used extensively as a model for cell biology of
apicomplexan organisms.
 advantages of using T. gondii for cell biology
 large enough to be easily seen under the
light microscope
 it can be grown in virtually any warm-
blooded cell Line
 only one species that infects all hosts
 Toxoplasmosis…
 Incidence
 varies greatly by country and by age
 1/3 of the global population
 congenital toxoplasmosis:1-10 /10,000 live births
 Immunocompromised individuals are at risk for
reactivation of latent infection, including potentially
fatal encephalitis.
 Causes and Risk Factors
 The house cat serves as definitive hosts

 Routes of transmission
• accidental ingestion of oocysts passed in cat
• ingesting tissue cysts
 raw or undercooked meat (lamb, pork and beef )
 drinking unpasteurized milk
 contaminated water, or unwashed fruits or
vegetables
• direct transmission of tachyzoites from mother to fetus
(congenital infection)
• blood transfusion /organ transplantation
Morphology
Tachyzoite-the
intracellular parasites,
3x6µm in size
Crescent or oval in
shaped, one end is
rounded and the other
end is pointed
Tachyzoites are the
actively proliferating
trophozoites, which are
observed during the acute
stage of infection
morphology…
Tissue cyst- filled with
bradyzoites
Bradyzoites crescent-shaped
cell and 7 x 1.5 μm measures
Bradyzoites in the chronic
stage of infection, develops
slowly and multiplies in the
tissues
Tissue cysts are found most
commonly in the brain and
in skeletal and cardiac
muscle
Cyst measures 10-100m
Morphology…

Oocysts: 

The oocyst contains

two sporocysts, each
of which contain four
sporozoites.   

10 µm in diameter
Life cycle
 complex
It involves three distinct inter-linked cycles
1. feline cycle
associate with formation of oocyst
oocyst can infect DH and IH
liberate sporozoites on reaching the intestinal
of cats
sporozoites penetrate the intestinal mucosal
cells
Life cycle…

Sporozoite multiply asexually(endopolygeny)

to form endozoites

After some cycle of multiplication,

gametogony is initiated → zygote is formed
→ covert to oocyst
Life cycle…
2. Feline- non feline cycle
IH infected by oocysts through contaminated
food and water
Liberate sporozoites on reaching the intestinal
of IH and multiply asexually to form endozoites
All cells except RBC can be infected
the infected cell rupture to release endozoites
Endozoites continue to multiply in different
sites
Endozoite convert to cystozoites that are
3. Non feline non feline cycle
Cystozoites in the tissue cysts of non-feline
animals are infective to other animals or
birds
Only asexaul multiplication takes place
Cystozoites infect the intestinal cells of non-
feline and intiate endopolgeny
They infect deeper tissues to form tissue
cysts containing cystozoite
 Cat ingests tissue cysts containing bradyzoites
 Gametocytes develop in the intestine
 Oocysts appear in the cat’s feces 3-5 days after
infection by cysts
 Oocysts require oxygen and they sporulate in 2-
3 days
Epidemiology and transmission

 Transmission to humans
 Oral
 Ingestion of under cooked Pork or Lamb
meat –tissue cyst.
 Exposure to oocysts
 Ingestion of contaminated food and water
 Direct contact with cat feces.
 Others
 Transplacental (Vertical transmission)
 Blood Product Transfusion
 Organ Transplantation
 The life cycle of Toxoplasma gondii begins
when a cat ingests toxoplasma-infected tissue
from an intermediate host(rodent)
 Tissue cysts within the muscle fibers or brain
are digested in the cat’s digestive tract.
 The parasite multiplies in the intestine of the cat
(sexual development) is eventually shed in cat
feces
Epidemiology:

 Human infection:
 Ingesting raw meat

 Sporulated oocysts from contaminated soil

 Birds & rodents are important in picking oocysts from soil or

scavenging bradizoites from infected animals

 WW distribution
 Hot, dry climates-lower incidence

 Temperate, humid climates-high incidence

Pathology
 Toxoplasma gondii invades numerous organs,
infecting a broad spectrum of cell types.
 Tachyzoites→macrophages→blood→organs(invasion
,asexual multiplication) → cell death.
 The resulting necrosis attracts inflammatory host
cells(lymphocytes and monocytes) → pathology.
 As host resistance develops, tissue cysts may form in
many organs(brain and muscle)-quiescent cysts
 evade the adaptive host immune
 tissue cysts periodically rupture, the released
bradyzoites are killed by the host immune system.
 If immunocompromised:bradyzoites develop
into tachyzoites- active toxoplasmosis
 3 categories
a. Postnatally acquired toxoplasmosis
b. Immunocompromised related
toxoplasmosis
c. Congenital toxoplasmosis
Postnatally acquired toxoplasmosis
 > 80-90% of primary infections- asymptomatic
 Mostly recognized finding
 cervical lymphadenopathy accompanied by low-grade

fever
 mononucleosis-like syndrome(Epstein-Barr virus)

 fever
 swelling of the lymph nodes
 hepatosplenomegally
 myalgia
 Atypical lymphocytosis in the blood
Enlarged cervical lymph nodes (arrows) in a 19-year-old boy
with acquired toxoplasmosis
Sever disease
 retinitis, which may result in strabismus(cross-eye) or
blindness.
 A characteristic residual pigmented scar is left on the
retina after resolution of infection.
 immunocompromised adults
 Brain lesions
 fever, headache, confusion & seizures
 Systemic manifestations
 Myocarditis & pneumonitis
 Immunocompromised toxoplasmosis
 Cerebral manifestations
 Altered mental status
 Focal Neurological deficit
 motor weakness
 speech disturbances
 cranial nerve palsy
 movement disorders
 visual defects
 sensory ,cerebelar dysfunction
Congenital toxoplasmosis
Congenital toxoplasmosis
 mother infected before conception-no congenital
toxoplasmosis
 new infections with detectable maternal parasitemia
 ~50% transmission rate of congenital toxoplasmosis
 transmission rate and severity of disease in the fetus are
inversely proportional
 mothers who develop acute toxoplasmosis in the first
trimester have a much lower fetal transmission rate
 fetuses exposed early are at much higher risk for severe
symptoms or death and spontaneous abortion
Gestational age at Risk of congenital Development of
maternal infection(95% CI) clinical signs in
seroconversion weeks % infected
offsprings(95% CI)%
13 6(3-9) (34-85)
26 40(33-47) 25(18-33)
36 72(60-81) 9(4-17)

Table 1. Risk of Toxoplasma gondii congenital infection

(transmission) and development of clinical signs in offspring
before age 3 years, according to gestational age at maternal
seroconversion
 Two types:
• Asymptomatic (inapparent) congenital toxo

 60% of infected
 may suffer from Long Term Sequelae
 ~90 percent of infected babies appear normal
at birth
 80 to 90 percent will develop sight-threatening
eye infections months to years after birth
 ~10 percent will develop hearing loss and/or
learning disabilities
 Symptomatic Congenital toxoplasmosis
 40% of infected
 About one in 10 infected babies has a severe
Toxoplasma infection that is evident at birth
 severe eye infections, an enlarged liver and
spleen, jaundice
 Some die within a few days of birth
 some suffer from mental retardation, severely
impaired eyesight and seizures
 Severe damage to fetus = stillbirth or abortion
 Hydrocephalus with bulging forehead (left) and
microophothalmia on the left eye
 Hydrocephalus
• rare symptom- increases in the volume of cerebrospinal fluid
Figure 1.Hydrocephalic and spastic baby

(CSF) within the ventricles of the brain as a result of host

reaction to the parasite
 Lab. Diagnosis
 Morphologic Dx
 tachyzoites in
 circulating WBC
 bone marrow
 lung
 spleen
 Histopathologic examination
 Culture or animal inoculation (rare)
 Serologic (mainly)
 Molecular Dx-
 Detection of T. gondii parasites in c.s.f.
• The c.s.f. usually contains neutrophils and
mononuclear cells.
 Method

1. Centrifuge the c.s.f. in a conical tube at medium

to high speed(about 1 000 g for 5 minutes).
o Transfer the supernatant fluid to another tube.

This can be used for protein testing (raised in

Toxoplasma encephalitis).
2. Tap the bottom of the tube to mix the sediment.
Spread a drop on a slide and allow the smear to air-
dry. Fix with methanol for 1–2 minutes.

3. Stain the smear using Giemsa technique

4. When dry examine the preparation microscopically

 40X objective to detect the parasites & 100X

objective to identify the tachyzoites.
 Serological diagnosis of toxoplasmosis
A. Sabin-Feldman dye test: the most reliable test for
the diagnosis of acute toxoplasmosis in pregnancy,
and to test neonates
• is highly sensitive and specific test
• It is performed in Reference Laboratories

B. The Eiken Toxoreagent latex agglutination test- is a

simpler test
• a quantitative test in microtitration plates
 the latex particles are coated with inactivated T. gondii
soluble antigen.
 A positive control is included in each test kit.
o Interpretation of test results
 Interpretation is difficult because of the high
prevalence of antibodies in most populations
due to past and subclinical infections
 detection of Toxoplasma specific IgM is
indicative of recent infection
 In diagnosing congenital toxoplasmosis
• + of IgM (which does not cross the placenta)
in the infant’s circulation is diagnostic
• Specific IgG in the infant’s circulation may be
of maternal origin or due to infection.
 Testing of the infant’s blood at 2 monthly

intervals will show whether the IgG antibody

level is decreasing

 At 6–10 months, the infant’s circulation should
not contain maternal Ig G
 persistence of Ig G beyond this time is
indicative of infection in the infant.
Serology……
 unreliable in immunodeficient (AIDS) pts

 normally IgM and IgG rise simultaneously

 IgG - persists for years

 IgM - undetectable after “cure”
 elevated IgM titer is diagnostic of recent infection

 A negative IgG or IgM test excludes Diagnosis

 both should be + if acutely infected

 If IgG screening is + then do IgM test
 a + IgM test confirms acute toxoplasmosis
or current Toxoplasma infection (measure
IgM antibodies, have low specificity and the
reported results are frequently
misinterpreted.
 In addition, IgM antibodies can persist for
months to more than one year.
 Submit IgG positive sera to reference
aboratory for IgM testing for diagnosis of
acute Toxo
 Treatment

 pyrimethamine + sulfadiazine

 Spiramycin-Pregnant
 Prevention:
 wear gloves when handling

soil
 hand washing after outdoor

activities
 cook all meats thoroughly
 keep cats